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    nrStar™ tRNA PCR芯片(H/M)

    • 簡介
    • tRNA研究热点
    • 芯片設計理念
    • 數據庫
    • 說明書和分析工具
    •        转运RNA(Transfer Ribonucleic Acid,tRNA)是生物体内含量最为丰富的短链非编码RNA分子之一。它携带并转运氨基酸,参与蛋白翻译,是连接mRNA与蛋白质的重要桥梁。尽管tRNA广泛存在于生物体内,但不同机体基因组对于特定密码子的偏好性不同,从而导致tRNA表达谱的差异。密码子偏好性影响翻译效率和精确性[1-3]。因此,tRNA表达谱的改变对多种细胞生理过程有着显著影响。细胞增殖[4]、分化[4, 5]和凋亡[6]等一系列生命活动都伴随着tRNA水平的变化。另外,许多疾病也显示出tRNA水平和分布的紊乱。系列研究表明,tRNA谱的紊乱和肿瘤[5, 7-14]、Ⅱ型糖尿病[15]、亨廷顿舞蹈症[16]、甲型H1N1流感[17]、天花[17]以及HIV感染[18]等疾病病理活动密切相关。因此,tRNA谱的研究对于深入了解人体生理及病理过程十分关键。
             PCR芯片是研究特定类群基因有效和可靠的手段之一。Arraystar nrStarTM tRNA PCR Arrays(H/M)可分别检测185个人或小鼠的tRNA,覆盖了GtRNAdb数据库中所有的细胞核反密码子isoacceptors (运载相同氨基酸但反密码子不同的tRNA互称为isoaccepetor),提供isoacceptor和isodecoder(反密码子相同但躯干序列不同的tRNA之间互称为isodecoder)两种层面上的表达检测。


      產品列表

      产品名称 货号 规格
      nrStaTM Human tRNA PCR Array V2.0 AS-NR-001H-1 384-well (2*192) / plate
      nrStarTM Human tRNA PCR Array V2.0 (Roche Light Cycler 480) AS-NR-001H-1-R 384-well (2*192) / plate
      nrStarTM Mouse tRNA PCR Array AS-NR-001M-1 384-well (2*192) / plate
      nrStarTM Mouse tRNA PCR Array (Roche Light Cycler 480) AS-NR-001M-1-R 384-well (2*192) / plate
      nrStarTM Human tRNA PCR Array V1.0 AS-NR-001-1 384-well (4*96) / plate


      優勢

      • 检测GtRNAdb数据库中全部的反密码子isoacceptor

      • 检测实时定量PCR可以区分的全部tRNA isodecoder

      • 聚焦可信度高的人tRNA (tRNAScan score >50)

      • 引物在多种类型组织和细胞中经过充分验证

      • 简单易操作,短时间内即可获得丰富数据结果


      为保证获得准确可靠的tRNA anti-codon isoacceptors和 isodecoders表达水平,推荐使用rtStarTM tRNA-optimized First-Strand cDNA Synthesis Kit (Cat# AS-FS-004) ,该试剂盒能高效去除tRNA上丰富的甲基化修饰,确保tRNA被准确检测。

    • tRNA的生物学功能,尤其是作为翻译调控分子发挥作用的文献综述选读

      1. Pan T: Modifications and functional genomics of human transfer RNA. Cell Res 2018.[PMID: 29463900]

      2. Schimmel P: The emerging complexity of the tRNA world: mammalian tRNAs beyond protein synthesis. Nat Rev Mol Cell Biol 2018, 19(1):45-58.[PMID: 28875994]

      3. Hanson G, Coller J: Codon optimality, bias and usage in translation and mRNA decay. Nat Rev Mol Cell Biol 2018, 19(1):20-30.[PMID: 29018283]

      4. Truitt ML, Ruggero D: New frontiers in translational control of the cancer genome. Nat Rev Cancer 2016, 16(5):288-304.[PMID: 27112207]

      5. Kirchner S, Ignatova Z: Emerging roles of tRNA in adaptive translation, signalling dynamics and disease. Nat Rev Genet 2015, 16(2):98-112.[PMID: 25534324]

      6. Suzuki T, Nagao A, Suzuki T: Human mitochondrial tRNAs: biogenesis, function, structural aspects, and diseases. Annu Rev Genet 2011, 45:299-329.[PMID: 21910628]



      tRNA isoacceptors 与 isodecoders的重要生理功能的原创文献选读

      1. Dittmar KA, Goodenbour JM, Pan T: Tissue-specific differences in human transfer RNA expression. PLoS Genet 2006, 2(12):e221.[PMID: 17194224]

      2. Pavon-Eternod M et al: tRNA over-expression in breast cancer and functional consequences. Nucleic Acids Res 2009, 37(21):7268-7280.[PMID: 19783824]

      3. Gingold H et al: A dual program for translation regulation in cellular proliferation and differentiation. Cell 2014, 158(6):1281-1292.[PMID: 25215487]

      4. Ishimura R et al: RNA function. Ribosome stalling induced by mutation of a CNS-specific tRNA causes neurodegeneration. Science 2014, 345(6195):455-459.[PMID: 25061210]

      5. Goodarzi H et al: Modulated Expression of Specific tRNAs Drives Gene Expression and Cancer Progression. Cell 2016, 165(6):1416-1427.[PMID: 27259150]

    • tRNA库的作用

      tRNA库,由运载每一个氨基酸的tRNA isoacceptor家族组成,在密码子使用、蛋白翻译效率和准确性、生物学过程及人类疾病研究中已成为重要的信息来源。密码子可以被单碱基错配的近似同源tRNA(near-cognate tRNA)解码。在tRNA库中,同源tRNA与近似同源tRNA的比率是被动态调节的,以影响翻译效率、保真度和转录本稳定性。在不同条件下,tRNA库可以通过多种方式发生变化以维持细胞稳态或促进某类基因的表达[1]。


      tRNA isodecoders的生物学重要性

      有相同反密码子但躯干序列不同的tRNA即为isodecoders。高度保守的tRNA isodecoder序列具有显著的组织表达特异性,且具有不同的功能。例如,小鼠 tRNA-Arg/UCU具有的5个isodecoder,其中一个isodecoder在中枢神经系统中特异性表达,具有维持神经细胞稳态和阻止神经退化的特殊功能(图1)。


      图.1. 小鼠 tRNA-Arg-TCT isodecoder 的cDNA 序列。与“管家”isodecoders相比,中枢神经系统特异性的tRNA isodecoder具有维持神经细胞稳态和阻止神经退化的特殊功能[2].


      參考文獻

      [1] Gavin H. et al. (2014) "Codon optimality, bias and usage in translation and mRNA decay." Molecular Cell Biology 19, 20-30 [PMID: 29018283]

      [2] Ishimura R. et al. (2014) "RNA function. Ribosome stalling induced by mutation of a CNS-specific tRNA causes neurodegeneration." Science 345(6195):455-9 [PMID: 25061210]

    • Human V2.0 tRNA List (GtRNAdb and mitotRNAdb)

      No. tRNA NAME tRNA ID
      1 Ala-AGC-1 Homo_sapiens_tRNA-Ala-AGC-1-1/ Homo_sapiens_tRNA-Ala-AGC-2-1~2/
      Homo_sapiens_tRNA-Ala-AGC-3-1/ Homo_sapiens_tRNA-Ala-AGC-4-1/
      Homo_sapiens_tRNA-Ala-AGC-5-1/ Homo_sapiens_tRNA-Ala-AGC-6-1/
      Homo_sapiens_tRNA-Ala-AGC-7-1/ Homo_sapiens_tRNA-Ala-AGC-8-1~2/
      Homo_sapiens_tRNA-Ala-AGC-10-1/ Homo_sapiens_tRNA-Ala-AGC-11-1/
      Homo_sapiens_tRNA-Ala-AGC-12-1~3/ Homo_sapiens_tRNA-Ala-AGC-13-1~2/
      Homo_sapiens_tRNA-Ala-AGC-14-1/ Homo_sapiens_tRNA-Ala-AGC-15-1
      2 Ala-AGC-2 Homo_sapiens_tRNA-Ala-AGC-3-1
      3 Ala-AGC-3 Homo_sapiens_tRNA-Ala-AGC-9-1~2
      4 Ala-AGC-4 Homo_sapiens_tRNA-Ala-AGC-10-1

      >>Full List


      Mouse tRNA List (GtRNAdb and mitotRNAdb)

      No. tRNA NAME tRNA ID
      1 Ala-AGC-1 Mus_musculus_tRNA-Ala-AGC-4-1~2/ Mus_musculus_tRNA-Ala-AGC-5-1~3/
      Mus_musculus_tRNA-Ala-AGC-6-1/ Mus_musculus_tRNA-Ala-AGC-7-1/
      Mus_musculus_tRNA-Ala-AGC-10-1
      2 Ala-AGC-2 Mus_musculus_tRNA-Ala-AGC-2-1~2
      3 Ala-AGC-3 Mus_musculus_tRNA-Ala-AGC-3-1
      4 Ala-AGC-4 Mus_musculus_tRNA-Ala-AGC-7-1
      5 Ala-AGC-5 Mus_musculus_tRNA-Ala-AGC-9-1
      6 Ala-AGC-6 Mus_musculus_tRNA-Ala-AGC-10-1
      7 Ala-AGC-7 Mus_musculus_tRNA-Ala-AGC-14-1

      >>Full List


      Human V1.0 tRNA List (GtRNAdb and mitotRNAdb)

      No. tRNA NAME tRNA ID
      1 Ala-AGC tdbD00003849/tdbD00003850
      2 Ala-CGC tdbD00003856/tdbD00003855
      3 Ala-GGC tRNA-Ala-GGC-chr19-3
      4 Ala-TGC tdbD00003861/tdbD00003862
      >>Full List



    • Aksomics使用Arraystar的芯片为客户提供服务,该产品说明书及分析工具见下:

      說明書:

      adobe_pdf_small Manual_nrStar™_Human_tRNA_PCR_Array_v2.0

      adobe_pdf_small Manual_nrStar™_Human_tRNA_PCR_Array_v1.051

      adobe_pdf_small Manual_nrStar™_Mouse_tRNA_PCR_Array_v1.01


      分析工具:

        excel Analysis_Tool_for_nrStar™_Human_tRNA_PCR_Array_v2.0

        excel Analysis tool for nrStar™ Human tRNA PCR Array - v1.01

        excel Analysis Tool for nrStar™ Mouse tRNA PCR Array_v1.0


      分析工具使用說明

        adobe_pdf_small Instructions of Human tRNA qPCR data analysis tool



      適用的qPCR仪:

      ABI ViiA™ 7

      ABI 7500 & ABI 7500 FAST

      ABI 7900HT

      ABI QuantStudio™ 6 Flex Real-Time PCR system

      ABI QuantStudio™ 7 Flex Real-Time PCR system

      ABI QuantStudio™ 12K Flex Real-Time PCR System

      Bio-Rad CFX384

      Bio-Rad iCycler & iQ Real-Time PCR Systems

      Eppendorf Realplex

      QIAGEN Rotor Gene Q 100

      Roche Light Cycler 480

      Stratagene Mx3000

      Roche Light Cycler 480